Pharmaceutical composition comprising glutarimide derivatives and use thereof in the treatment of eosinophilic diseases

ABSTRACT

The present invention relates to novel biologically active glutarimide derivatives of general formula (I) or a pharmaceutically acceptable salt thereof, their use as a therapeutic agent for the treatment of eosinophilic diseases, preferably of allergic nature, in particular bronchial asthma, allergic rhinitis, polypous rhinosinusopathies, eosinophilic colitis, eosinophilic syndrome, allergic conjunctivitis, atopic dermatitis, Churg-Strauss syndrome, anaphylactic shock, Quincke&#39;s edema, eosinophilic vasculitis, eosinophilic esophagitis, eosinophilic gastroenteritis, or fibroses. The invention also relates to pharmaceutical compositions comprising glutarimide derivatives of general formula (I):

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.15/036,603, now pending, which is a U.S. National Stage applicationunder 35 U.S.C. § 371 of International Application PCT/RU2014/000855(published as WO 2015/072893 A1), filed Nov. 12, 2014, which claimspriority to Application RU 2013150861, filed Nov. 14, 2013. Benefit ofthe filing date of each of these prior applications is hereby claimed.Each of these prior applications is hereby incorporated by reference inits entirety.

FIELD OF THE INVENTION

The invention relates to the use of biologically active glutarimidederivatives or pharmaceutically acceptable salts thereof as agents forthe treatment of eosinophilic diseases.

BACKGROUND

Eosinophils are cells of the innate immunity. They are produced in thebone marrow and preferably circulate in the blood. The main effectorfunction of eosinophils is an immediate release of cytoplasmic granulesin response to activation by various stimuli. Cytoplasmic granulescomprise pro-inflammatory mediators: cytokines, chemokines, lipid- andneuromediators, growth factors, and cationic proteins. The cationicproteins of eosinophils include 4 classes: major basic protein (MBP),eosinophil peroxidase (EPO), eosinophil cationic protein (ECP), andeosinophil-derived neurotoxin (EDN). In combination, these proteins havea cytotoxic action on both infectious microorganisms and tissues of ahost, causing eosinophilic inflammation [Hogan S P, Rosenberg H F,Moqbel R, et al. Eosinophils: biological properties and role in healthand disease//Clin Exp Allergy. 2008; 38(5):709-50]. The eosinophil countin the blood is normally 0.02-0.3 10⁹/L, or 0.5 to 5% of the totalleukocytes. An increased blood eosinophil count relative to the normallevel is eosinophilia. Hypereosinophilia or large eosinophilia is acondition when the content of eosinophils in the blood is 15% or more,usually when the total leukocyte count is increased [Hoffman R, Benz Jr.E J, Shattil S J, et al., eds. Hematology: Basic Principles andPractice. 4th ed. Philadelphia, Pa.: Churchill Livingston; 2005: 768].

An increased level of eosinophils in the blood and tissues accompaniesdiseases of various etiology and pathogenesis. They include parasiticinvasions, a broad spectrum of allergic diseases, such as asthma,rhinitis, nasal polyps, eosinophilic colitis, eosinophilic syndrome,allergic conjunctivitis, and atopic dermatitis; rheumatic diseases(rheumatoid arthritis, diffuse eosinophilic fasciitis, Churg-Strausssyndrome, nodular periarteritis); and pathologies of unclear etiology(eosinophilic esophagitis, eosinophilic gastroenteritis) [Blanchard C,Rothenberg M E. Biology of the eosinophil// Adv Immunol. 2009;101:81-121].

Among allergic diseases, bronchial asthma, which is a chronicinflammatory airway disease that is characterized by episodic airflowobstruction, inflammation of the respiratory tract, and by an increasedbronchial reactivity to non-specific allergens, is medically mostimportant.

There are a lot of evidences that eosinophils are a key component of anallergic response in asthma. IL-3 and IL-5 secreted by mast cellsprovide the accumulation of eosinophils in lungs, followed by activationof these cells, which is accompanied by the release of LTC4, eosinophilcationic protein, major basic protein, neurotoxin, eosinophil peroxidase(EPO), transforming growth factor, and free radicals [Blanchard C,Rothenberg M E. Biology of the eosinophil// Adv Immunol. 2009;101:81-121].

The activity of the inflammatory process in asthma has been found to bein direct correlation with the serum level of eosinophil cationicprotein [Bjornsson E., Janson C., Hakansson L. et al. Serum eosinophilcationic protein in relation to bronchial asthma in young Swedishpopulation. Allergy 1994; Vol. 49: 400-407]. Lavage fluid of patientswith bronchial asthma has an increased eosinophil count. The eosinophilcell surface has low-affinity receptors for IgE and due to thateosinophils may be directly activated by cause-significant allergens.The eosinophil cell surface has further been found to have receptors forIL-2, IL-3, IL-5, GM-CSF, PAF, and prostaglandins. Through thesereceptors, the above-mentioned cytokines and lipid mediators are able toinduce the activation of eosinphils that release mediators (LTC4, PAF)and cytokines (IL-3, IL-4, IL-5, IL-8, GM-CSF, TGFβ). Destruction of theairway epithelium, which results from the action of eosinophil proteins,causes the development of bronchial hyperreactivity and a reduction inthe barrier function of the airway epithelium.

Now, a great attention is being attached to the role of eosinophils inthe regeneration and remodulation of tissues because of a clearrelationship that have been found to exist between eosinophilia intissues and some fibrous diseases (endomyocardial fibrosis complicatedwith hepatic fibrosis in patients with hypereosinophilic syndrome,nodular sclerosing Hodgkin's disease, and subepithelial fibrosis inbronchial asthma) [Noguchi H. et al. Tissue eosinophilia and eosinophildegranulation in syndromes associated with fibrosis// Am. J. Pathol.1992, Vol. 140. P. 521-528].

Eosinophils are a source of a number of fibrogenic and growth factors,including transforming growth factor-β(TGF-β) fibroblast growth factor(FGF)-2, vascular endothelial growth factor (VEGF), matrixmetalloproteinase (MMP)-9, IL-1β, IL-13, and IL-17. Clinical trialsinvolving anti-IL-5 antibodies also supported the role of eosinophils inthe events associated with the deposition of specific matrix proteins inthe reticular basement membrane [Kay A B, Phipps S, Robinson D S. A rolefor eosinophils in airway remodeling in asthma//Trends Immunol. 2004,Vol. 25, P. 477-82].

Today, the most common method for treating asthma is the use ofcorticosteroids (budesonide, beclomethasone dipropionate, fluticasonepropionate, mometasone furoate) by inhalation. However, corticosteroidsfunction by inducing a general immunosuppressive action, and there areadverse side effects caused by long-term administration thereof, such ashigh blood pressure, osteoporosis, and cataract development [Barnes P J.New drugs for asthma//Semin Respir Crit Care Med. 2012; 33(6):685-94].Corticosteroids should be administered every day, and patient'scompliance with this requirement is therefore another problem for thesuccessful use of said therapeutic agents. In addition, there arecorticosteroid-insensitive patients who need an alternative therapy.Selective targeting to eosinophils may overcome the side effects causedby the use of systemic immunosuppressive agents with pleiotropic action.

The drugs for reducing eosinophilia by inhibiting the interactionbetween interleukin-5 and receptor IL-5Rα on the eosinophil cell surfaceare currently in clinical trials. Such drugs include humanizedmonoclonal anti-IL-5 antibodies (mAt) and concurrent IL-5Rα inhibitors.

Among IL-5 neutralizing monoclonal antibodies, SB240563 (mepolizumab,Glaxo Smith Kline) is most effective. There are reports [Nair P,Pizzichini M M M, Kjarsgaard M, et al. Mepolizumab forprednisone-dependent asthma with sputum eosinophilia. NEJM. 2009;360:985-93] that mepolizumab therapy of patients withprednisolone-dependent asthma reduces eosinophilia in the blood andsputum and, most importantly, improves patient's quality by reducingexacerbation frequency and a prednisolone dose. Another clinical trialshowed that the administration of anti-IL-5 antibodies (mepolizumab) tothe patients with corticosteroid-insensitive asthma also led to areduction in exacerbation frequency and improved patient's qualityaccording to the AQLQ (Asthma Quality of Life Questionnaire). Inaddition to the action on asthma, this trial also showed a therapeuticeffect against polypous rhinosinusopathy [Haldar P, Brightling C E,Hargadon B, et al. Mepolizumab and exacerbations of refractoryeosinophilic asthma. NEJM. 2009; 360:973-84].

In an independent clinical trial in adults with polypousrhinosinusopathy, mepolizumab significantly reduced the levels of ECPand a soluble form of IL-5Rα in the blood, and the concentration ofIL-5Rα, IL-6, and IL-1b in the nose, which correlated with analleviation of the disease, according to the total polyp score [GevaertP, Van Bruaene N, Cattaert T, et al. Mepolizumab, a humanized anti-IL-5mAb, as a treatment option for severe nasal polyposis. J Allergy ClinImmunol. 2011; 128(5):989-995].

The use of anti-IL-5 antibodies is not limited to bronchial asthma andpolypous rhinosinusopathy. This therapy is also effective in othereosinophil-mediated diseases. For example, in patients withhypereosinophilic syndrome, mepolizumab therapy reduced eosinophilia inthe blood and made it possible to reduce the administered dose ofprednisolone [Rothenberg M E, Klion A D, Roufosse F E, et al. Treatmentof patients with the hypereeosinophilic syndrome with mepolizumab. NEJM.2008; 358(12):1215-28]. Patients with eosinophilic esophagitis treatedwith anti-IL-5 antibodies showed an improved clinical picture associatedwith a reduced dysphagia and a six-fold reduction in the bloodeosinophil count, and in some patients, esophageal epithelialhyperplasia was reduced [Stein M L, Collins M H, Villanueva J M, et al.Anti-IL-5 (mepolizumab) therapy for eosinophilic esophagitis. J AllergyClin Immunol. 2006; 118(6):1312-9]. Clinical trials of the drug inchildren showed that the patients having in the blood not more than 20eosinophils in one field of microscope had improved symptoms, such asredness, fragility, and grooves and vertical lines on the esophagealmucosa [Assa'ad A H, Gupta S K, Collins M H, et al. An antibody againstIL-5 reduces numbers of esophageal intraepithelial eosinophils inchildren with eosinophilic esophagitis. Gastroenterology. 2011;141(5):1593-604].

Mepolizumab is also successfully used as therapy of eosinophilicvasculitis [Kahn J E, Grandpeix-Guyodo C, Marroun I, et al. Sustainedresponse to mepolizumab in refractory Churg-Strauss syndrome. J AllergyClin Immunol. 2010; 125:267-70]. In a 28-aged female, monthlyadministration of mepolizumab reduced the blood eosinophil count tonormal, prevented the formation of corticosteroid asthma, and based onx-ray data, improved the condition of the lung parenchyma [Kim S,Marigowda G, Oren E, Israel E, Wechsler M. Mepolizumab as asteroid-sparing treatment option in patients with Churg-Strausssyndrome. J Allergy Clin Immunol. 2010; 125:1336-43]. In clinical trialsin patients with eosinophilic vasculitis and pronounced eosinophilia,mepolizumab therapy made it possible to reduce the dose ofcorticosteroids. Eosinophilia also reduced, but after the trialcompletion, the exacerbation was repeated [Oldhoff J M, Darsow U, WerfelT, et al. Anti-IL-5 recombinant humanized monoclonal antibody(mepolizumab) for the treatment of atopic dermatitis. Allergy. 2005;60(5):693-6].

Mepolizumab is reported [Amini-Vaughan Z J, Martinez-Moczygemba M,Huston D P. Therapeutic strategies for harnessing human eosinophils inallergic inflammation, hypereosinophilic disorders, and cancer //CurrAllergy Asthma Rep. 2012, Vol. 12, No 5. P. 402-412] hat to be in thesecond phase of clinical trials of therapy for asthma, eosinophilicesophagitis in adults, eosinophilic esophagitis in children,eosinophilic vasculitis, and polypous rhinosinusopathies, and in thethird phase of clinical trials as a treatment of hypereosinophilicsyndrome, eosinophilic esophagitis in children, rhinovirus-inducedasthma, and chronic obstructive bronchitis. Another medicament,reslizumab (Cephalon), which is also a humanized monoclonal anti IL-5antibody SCH55700, is in the second phase of clinical trials as atherapy for hypereosinophilic syndrome and loiasis, and in the thirdphase as a therapy of asthma and eosinophilic esophagitis in children.All these allow for concluding that selective therapy aimed at reducingeosinophilia is a perspective approach to the treatment of diseasesmediated by this cell type (bronchial asthma, allergic rhinitis,allergic conjunctivitis, atopic dermatitis, polypous rhinosinusopathy,eosinophilic esophagitis, eosinophilic vasculitis, and hypereosinophilicsyndrome).

However, mAt therapy has several drawbacks. Monoclonal antibodies areexpensive therapeutic agents that should be administered for a month ortwo. An important factor is a problem of patient's non-compliance withphysician's order, which is because of multiple visits to a physician'soffice to receive the drug injections. In addition, allotype divergencebetween a patient and a therapeutic antibody can lead to that themonoclonal antibody-based therapy becomes ineffective. A high dose ofmAt and a possibility of forming immune complexes also may reduce theefficiency of passive immunization.

Other methods providing therapeutic agents against pathologicalconditions characterized by eosinophilia are disclosed in WO 97/45448and WO 03/040164. Application WO 97/45448 provides the use of “modifiedand variant forms of IL5 molecules capable of antagonizing or reducing,in another way, the activity of IL-5” to improve, alleviate, or reduceeffects deviated from the norm, which are caused by native and mutantforms of IL5. It is reported that the antagonizing action is a result ofvariant forms of IL5 that bind with the low-affinity chain of IL5R butnot with high-affinity receptors. Acting in such a way, the variantscompete with IL5 for binding with its receptors without any effect onthe physiological action of IL5.

Application WO 03/040164 provides a composition for vaccination directedto endogenous formation of antibodies to IL-5, IL-13, and eotoxin, i.e.to key factors of maturation, activation, localization, and vitality ofeosinophils. The composition comprises a virus-like particle and atleast one protein or peptide of IL-5, IL-13 and/or eotaxin boundthereto. According to the invention, said composition is useful in theproduction of vaccines for the treatment of allergic diseases with aneosinophilic component and as a pharmaccine to prevent or cure allergicdiseases with an eosinophilic component.

Application Ser. No. 8501176 provides the use of antibodies binding toIL-5R. These antibodies comprise a binding site recognizing IL-5receptors (IL-5R) and Fc-fragment. The claimed method reduces theeosinophil count in the blood, marrowbone, gastro-intestinal tract (forexample, esophagus, stomach, small intestine and large gut), or lungs,thereby reducing clinical manifestations of asthma and chronicobstructive bronchitis of lungs in human beings(http://www.patentgenius.com/patent/8501176.html).

Yong Sup Lee et al. in Studies on the site-selective N-acyliminium ioncyclazation: synthesis of (±)-glochidine and (±)-glochidicine,Heterocycles, Vol 37, No 1. 1994, disclose the preparation of histaminesuccinimide by fusing histamine dihydrochloride together with succinicanhydride under heating the initial reagents to 200-230° C. for 40minutes.

The publication of international application WO 2007/007054 disclosessuccinimide and glutarimide derivatives of general formula (I), havinginhibitory action on DNA methylation in cells, in particular tumorcells. Compounds disclosed in said publication are prepared by anaddition reaction between an amino derivative comprising a hydrocarbonchain and a corresponding anhydride or acid, or ester, followed byoptional cyclization, if necessary in the presence of a base.

The described methods of synthesis imides of glutaric acid compriseheating a dicarboxylic acid or a derivative thereof, such as anhydride,diester, etc., with a primary amine or amide thereof (thermalcyclization) [Weigand-Hilgetag, Eksperimentalnye metody v organicheskoikhimii [Experimental Methods in Organic Chemistry], ed. by N. N.Suvorov, M., Khimiya, 1968; p. 446], cyclization of monoamides ofcorresponding dicarboxylic acids by using a dehydrating agent as acarboxylic group-activating reagent, such as acetic anhydride [Shimotoriet al, Asymmetric synthesis of δ-lactones with lipase catalyst. Flavourand Fragrance Journal, 2007, V. 22, No. 6, P. 531-539], acetyl chloride[Ito et al., Chemoselective Hydrogenation of Imides Catalyzed byCpRu(PN) Complexes and Its Application to the Asymmetric Synthesis ofParoxetine.//Journal of the American Chemical Society, 2007, V. 129, No.2, P. 290-291], carbonyldiimidazole [Polniaszek, et al., Stereoselectivenucleophilic additions to the carbon-nitrogen double bond. 3. Chiralacyliminium ions.//Journal of Organic Chemistry, 1990, V. 55, No. 1, P.215-223], glutaric or succinic anhydrides [Ainhoa Ardeo et al, Apractical approach to the fused β-carboline system. Asymmetric synthesisof indolo[2,3-α]indolizidinones via a diastereoselective intramolecularα-amidoalkylation reaction. /Tetrahedron Letters, 2003, 44, 8445-8448].

The international publication of patent application WO2007/000246provides a method of synthesis of glutarimides by alkylation ofpiperidine-2,6-dione and pyrrolidin-2,5-dione with corresponding haloderivatives in DMF, followed by separating the target substituted imidederivatives by preparative chromatography, which is not applicable forthe synthesis of macro amounts.

Thus, the object of the present invention is the use of non-toxicglutarimide derivatives effective for the treatment of eosinophilicdiseases, preferably of allergic nature, such as bronchial asthma,allergic rhinitis, polypous rhinosinusopathies, eosinophilic colitis,eosinophilic syndrome, allergic conjunctivitis, atopic dermatitis,Churg-Strauss syndrome, anaphylactic shock, Quincke's edema,eosinophilic vasculitis, eosinophilic esophagitis, eosinophilicgastroenteritis, and fibroses.

SUMMARY OF THE INVENTION

The present invention relates to use of glutarimide derivatives ofgeneral formula (I):

wherein R₁ and R′₁ are independently hydrogen or C₁-C₆alkyl, forexample, methyl;

R₂ is

optionally substituted with C₁-C₆alkyl, for the treatment ofeosinophilic diseases, preferably of allergic nature, such as bronchialasthma, allergic rhinitis, polypous rhinosinusopathies, eosinophiliccolitis, eosinophilic syndrome, allergic conjunctivitis, atopicdermatitis, Churg-Strauss syndrome, anaphylactic shock, Quincke's edema,eosinophilic vasculitis, eosinophilic esophagitis, eosinophilicgastroenteritis, and fibroses, as disclosed in application RU 2013116826of Dec. 4, 2013.

The inventors have found that glutarimide derivatives suppresseosinophilia in various inflammation models in all tested media (blood,bronchoalveolar lavage (BAL), and tissues). In particular, in the modelof sephadex-induced lung inflammation in rats, glutarimide derivativesreduced the eosinophil count in BAL, and in the model ofovalbumin-induced asthma in guinea pigs, glutarimide derivatives reducedeosinophilia in BAL and blood.

Thus, the present invention relates to a therapeutic method for treatingeosinophilic diseases, preferably of allergic nature, such as bronchialasthma, allergic rhinitis, polypous rhinosinusopathies, eosinophiliccolitis, eosinophilic syndrome, allergic conjunctivitis, atopicdermatitis, Churg-Strauss syndrome, anaphylactic shock, Quincke's edema,eosinophilic vasculitis, eosinophilic esophagitis, eosinophilicgastroenteritis, and fibroses, the method comprising administering to apatient an effective amount of a glutarimide derivative of generalformula (I) or a pharmaceutically acceptable salt thereof.

The present invention also relates to a therapeutic agent for thetreatment of eosinophilic diseases, preferably of allergic nature, suchas bronchial asthma, allergic rhinitis, polypous rhinosinusopathies,eosinophilic colitis, eosinophilic syndrom, allergic conjunctivitis,atopic dermatitis, Churg-Strauss syndrome, anaphylactic shock, Quincke'sedema, eosinophilic vasculitis, eosinophilic esophagitis, eosinophilicgastroenteritis, and fibrosis, wherein the therapeutic agent is aglutarimide derivative of general formula (I) or a pharmaceuticallyacceptable salt thereof.

Another subject matter of the present invention is a pharmaceuticalcomposition for the treatment of eosinophilic diseases, preferably ofallergic nature, such as bronchial asthma, allergic rhinitis, polypousrhinosinusopathies, eosinophilic colitis, eosinophylic syndrome,allergic conjunctivitis, atopic dermatitis, Churg-Strauss syndrome,anaphylactic shock, Quincke's edema, eosinophilic vasculitis,eosinophilic esophagitis, eosinophilic gastroenteritis, and fibrosis,the composition comprising an effective amount of a glutarimidederivative of general formula (I) or a pharmaceutically acceptable saltthereof, and a pharmaceutically acceptable carrier.

The synthesis of the above-mentioned glutarimide derivative of generalformula (I) is disclosed in application RU 2013116826 of Dec. 4, 2013.

Compound used in the present invention may be prepared by a methodcomprising heating of initial dicarboxylic acid monoamides with adehydrating agent in an organic solvent or in the dehydrating agent assuch, optionally adding sodium acetate. The dehydrating agent used inthe method may include dicarboxylic acid anhydrides, organic acidchloroanhydrides, and carbonyldiimidazole.

Initial dicarboxylic acid monoamides and methods for preparing thereofare disclosed in the publication of international application WO1999/001103.

DETAILED DESCRIPTION OF THE INVENTION

Preferred compounds used in the present invention are compounds ofgeneral formula (I)

wherein R₁ and R′₁ are independently hydrogen or methyl;

R₂ is

The most preferred compounds of the present invention are compoundspresented in Table 1.

TABLE 1 The number of compound Structure 1

2

3

4

5

6

7

8

The pharmaceutically acceptable salts of the compounds according to thepresent invention may include additive salts of organic acids (forexample, formiate, acetate, maleate, tartrate, methanesulfonate,benzenesulfonate, toluenesulfonate, etc.), additive salts of inorganicacids (for example, hydrochloride, hydrobromide, sulphate, phosphate,etc.), and salts with amino acids (for example, an asparaginic acidsalt, a glutamic acid salt, etc.), preferably chlorohydrates andacetates.

Glutarimide derivatives of general formula (I) are therapeuticallyactive against eosinophilic diseases.

The compounds of the present invention may be used, in particular, forthe treatment of bronchial asthma, allergic rhinitis, polypousrhinosinusopathies, eosinophilic colitis, eosinophilic syndrome,allergic conjunctivitis, atopic dermatitis, Churg-Strauss syndrome,anaphylactic shock, Quincke's edema, eosinophilic vasculitis,eosinophilic esophagitis, eosinophilic gastroenteritis, and fibroses.

The compounds according to the present invention are administered in aneffective amount that provides a desired therapeutic effect.

The compounds of general formula (I) may be administered orally,topically, parenterally, intranasally, by inhalation, and rectally in aunit dosage form comprising a non-toxic pharmaceutically acceptablecarrier.

The term “parenteral administration” as used herein means subcutaneous,intravenous, intramuscular injection, or infusion.

The compounds according to the present invention may be administered toa patient at a dose of from 0.1 to 30 mg/kg of body weight once daily,preferably at a dose of from 0.25 to 10 mg/kg one or more times a day.

In addition, it should be noted that a particular dose for a particularpatient depends on many factors, including the activity of a usedcompound, patient's age, body weight, gender, general health condition,diet, and also on the time and route of administration of a therapeuticagent, the rate of its excretion from the body, a particularly usedcombination of therapeutic agents, and the disease severity in anindividual to be treated.

The pharmaceutical composition according to the present inventioncomprises a compound of general formula (I) in an amount effective toachieve a desired technical result, and may be administered in a unitedosage form (for example, solid, semi-solid, or liquid form) comprisingthe compound according to the present invention as an active ingredientin a mixture with a carrier or an excipient suitable for intramuscular,intravenous, oral, sublingual, inhalation, intranasal, and intrarectaladministration. In the composition, the active ingredient may be incombination with conventional nontoxic pharmaceutically acceptablecarriers suitable for the manufacture of solutions, tablets, pills,capsules, dragee, suppositories, emulsions, suspensions, ointments,gels, and any other dosage forms.

The compounds that can be used as an excipient are various compounds,such as saccharides, for example, glucose, lactose, or sucrose,mannitol, or sorbitol, cellulose derivatives, and/or calcium phosphate,for example, tricalcium phosphate or acidic calcium phosphate. Thecompounds that can be used as a binder, tare compounds such as starchpaste, for example, corn, wheat, rice, and potato starch, gelatin,tragacanth, methylcellulose, hydroxypropyl methylcellulose,carboxymethylcellulose, and/or polyvinylpyrrolidone. If necessary, theremay be used a disintegrating agent, such as the aforementioned starchesand carboxymethyl starch, crosslinked polyvinylpyrrolidone, agar, oralginic acid or a salt thereof, such as sodium alginate.

Additives that may be optionally used include flowability-control agentsand lubricants, such as silicon dioxide, talc, stearic acid and saltsthereof, such as magnesium stearate or calcium stearate, and/orpropylene glycol.

The core of a coted pill is usually coated with a layer that isresistant to gastric acid. A concentrated solution of saccharides thatmay optionally comprise gum arabic, talc, polyvinylpyrrolidone,polyethylene glycol, and/or titanium dioxide, and suitable organicsolvents or salts thereof may be used for this purpose.

Additives may also include stabilizers, thickening agents, colorants,and fragrances.

As an ointment base, there may be used hydrocarbon ointment bases, suchas white Vaseline and yellow Vaseline (Vaselinum album and Vaselinumflavum), Vaseline oil (Oleum Vaselini), and white ointment and liquidointment (Unguentum album and Unguentum flavum), and the substances,such as solid paraffin or wax, can be used as an additive providing afirmer texture; absorptive ointment bases, such as hydrophilic Vaseline(Vaselinum hydrophylicum), lanoline (Lanolinum), and cold cream(Unguentum leniens); water-removable ointment bases, such as hydrophilicointment (Unguentum hydrophylum); water-soluble ointment bases, such aspolyethylene glycol ointment (Unguentum Glycolis Polyaethyleni);bentonite bases; and others.

Methylcellulose, carboxymethylcellulose sodium salt, oxypropylcellulose,polyethylene glycol, polyethylene oxide, or carbopol may be used as abase for gels.

Bases for suppositories may be water-insoluble bases, such as cocoabutter; water-soluble or water-miscible bases, such as gelatin-glycerolor polyethylene oxide bases; or combined bases, such as soap-glycerolbases.

When preparing a unit dosage form, the amount of an active agent used incombination with a carrier may vary depending on a recipient under thetreatment and on a particular method of administration of thetherapeutic agent.

For example, when the compounds according to the present invention areused in the form of a solution for injections, the amount of the activeagent in this solution is up to 5 wt. %. A diluent may be a 0.9% sodiumchloride solution, distilled water, a Novocain solution for injection,Ringer's solution, a glucose solution, or a specific solubilizingadjuvant. When the compounds according to the present invention areadministered in the form of a tablet or suppository, their amount is upto 200 mg per unit dosage form.

Dosage forms according to the present invention are prepared byconventional procedures, such as blending, granulation, formation of acoating pill, dissolution, and lyophilization.

It should be noted that the compounds according to the present inventionhave no side effects and contraindications for administration. Inaddition, fatal cases in the experimental animals were not registered intoxicity tests of the compounds according to the present invention,administered orally at a dose of 1500 mg/kg.

The detailed description of the compounds according to the presentinvention, and studies of their pharmacological activity are disclosedin the following examples that are intended for purposes of illustrationonly and are not intended to limit the scope of the invention.

EXPERIMENTAL PART

The synthesis of the above-mentioned glutarimide derivative of generalformula (I) is disclosed in application RU 2013116826 of Dec. 4, 2013.

Examples of the Synthesis of Glutarimide Derivatives of General Formula(I)

Materials and Methods

The identity of the obtained compounds were assessed by the thin-layerchromatography (TLC) method on plates “Kieselgel 60 F254” (“Merck”,German) in a solvent system: chloroform-methanol (9:1) (1),chloroform-methanol (1:1) (2).

Chromatograms and electrophoregrams were stained with achlorotetramethylbenzene reagent and Pauly's reagent.

LC/MS system for analysis of multicomponent mixtures Shimadzu AnalyticalHPLC SCL10Avp; mass spectrometer PE SCIEX API 165 (150) (Canada).Conditions: column: Waters ACQUITY UPLC BEH C18 2.1×50 mm 1.7 μm,gradient elution system: water with 0.1% HCOOH - acetonitrile with 0.1%HCOOH.

Analytical-scale reversed phase HPLC was performed on a Shimadzu HPLCchromatograph under the following conditions: column: Luna C18(2) 100 A,250×4.6 mm (Serial number 599779-23), gradient elution system: aphosphate buffer solution (pH 3.0):methanol (condition A); column:Merk.LiChroCART 250×4 mm 5 μm LiChrospher 100RP-8E 5 μmC8 (Serial number1.50837.0001), gradient elution system: an ammonium acetate buffersolution (pH 7.5): acetonitrile (condition B); gradient elution system:a buffer containing 0.0025M sodium 1-hexylsulphonate (pH 3):acetonitrile(condition C); and column: Symmetry C18 150×4.6 mm, gradient elutionsystem: a buffer solution containing 0.0025M sodium 1-hexylsulphonate(pH 3):acetonitrile (condition D).

¹H NMR spectra were registered on a spectrometer (Bruker DPX-400,German).

High-resolution mass-spectra were obtained on a time-of-flight massspectrometer by a method of matrix-assisted laser-desorption ionizationwith 2,5-dihydroxybensoic acid used as a matrix, on an Ultraflex massspectrometer (“Bruker”, German).

Example 1 1-(2-(1H-imidazol-4-yl)ethyl)piperidine-2,6-dione (compound 1)

N,N′-dimethylformamide (60 mL) and 2-(imidazol-4-yl)-ethanamide ofpentandioic-1,5 acid (20 g) were filled in a 250 mL flat-bottom flask.Carbonyldiimidazole (17.3 g; 1.2 equiv.) was added under vigorousstirring. The reaction mixture was heated under stirring to 90° C. for 2hours. The reaction was controlled by ¹H-NMR spectroscopy (a sample (0.5mL) was diluted with a sulphuric ether, and the precipitate wasdissolved in DMSO-d₆). When the initial 2-(imidazol-4-yl)-ethanamide ofpentandioic-1,5 acid was absent in the reaction mass, the mass wascooled and poured out into a three-fold volume of methyl tert-butylether (180 mL). The reaction mixture was allowed to stand for 1 hour,and the precipitate was filtered, washed with 60 mL of methyl tert-butylether, and dried. The yield of the crude1-(2-(1H-imidazol-4-yl)ethyl)piperidine-2,6-dione was 12.4 g (67%).

The crude 1-(2-(1H-imidazol-4-yl)ethyl)piperidine-2,6-dione (12 g) andisopropanol (36 mg) were filled in a 100 mL flat-bottom flask. Themixture was heated to complete dissolution of the residue, then 1.2 g ofactivated carbon were added thereto, and the mixture was aged at boilingtemperature for an hour. The solution being hot was filtered through apre-heated ceramic filter. The residue on the filter was washed with 6mL of hot isopropanol. The hot stock solution was cooled to roomtemperature and allowed to stand for crystallization over night understirring. Precipitated crystals were filtered, washed with mL of coolisopropanol, and dried. After recrystallization, the amount of theobtained 1-(2-(1H-imidazol-4-yl)ethyl)piperidine-2,6-dione was 10.1 g(84%). Rf 0.43 (1). The product was analyzed with an LC/MS method: anindividual peak at a retention time of 1.57 min; [M+H]⁺=208. HPLC undercondition A: an individual peak at a retention time of 15.5 min. ¹H NMR(400.13 MHz, DMSO-d₆, δ, m.d., J/Hz): 1.81 (quint, 2H, CH₂CH₂CH₂, J=6.5Hz), 2.58 (m, 6H, CH₂C, CH₂CH₂CH₂); 3.82 (t, 2H, CH₂N, J=7.8 Hz), 6.77(br.s, 2H, CCH), 7.49 (s, 1H, NCHN), 11.81 (br.s, 1H, NH).

If necessary, in the synthesis of the compounds according to the presentinvention, the nitrogen atom in heterocycles may be protected by using,for example, a carbamate protecting group, such as tert-butoxycarbonylgroup (Boc).

The compounds presented in Table 2 were obtained according to theabove-indicated method.

TABLE 2 The number of a compound Structural formula Physical andchemical data 2

LC/MS: an individual peak at a retention time of 0.41 min, [M + H] ⁺ =208. HPLC under condition B: an individual peak at a retention time of16.72 min. ¹H-NMR (400.13 MHz, DMSO-d₆, δ, m.d., J/Hz): 1.82 (quint, 2H,CH₂CH₂CH₂, J = 6.5 Hz); 2.57 (t, 4H, CH₂CH₂CH₂, J = 6.5 Hz); 2.72 (t,2H, CH₂C, J = 7.5 Hz); 3.90 (t, 2H, CH₂N, J = 7.5 Hz); 6.86 (s, 2H,CH₂N, J = 7.5 Hz); 11.72 (br.s, 1H, NH) 3

LC/MS: an individual peak at a retention time of 0.41 min, [M + H] ⁺ =236. HPLC under condition A: an individual peak at a retention time of22.16 min. ¹H-NMR (400.13 MHz, DMSO-d₆, δ, m.d., J/Hz): 0.91 (s, 6H,CH₃); 2.58 (m, 6H, CH₂C, CH₂CCH₂); 3.86 (t, 2H, CH₂N, J = 7.3 Hz); 6.60,6.85 (br.s, 1H, CCH); 7.50 (br.s, 1H, NCHN); 11.8 (br.s, 1H, NH) 7

LC/MS: an individual peak at a retention time of 0.74 min, [M + H] ⁺ =211. HPLC under condition C, an individual peak at a retention time of10.8 min.¹H-NMR (400.13 MHz, DMSO-d₆, δ, m.d., J/Hz): 1.52 (m, 1H,pyrrolidine), 1.72 (m, 1H, pyrrolidine), 1.86 (m, 5H, pyrrolidine + CH₂CH + COCH₂CH₂ CH₂CO), 2.10 (m, 1H, pyrrolidine), 2.61 (t, 4H, COCH₂CH₂CH₂ CO, J = 6.4 Hz), 3.10 (m, 2H, pyrrolidine), 3.30 (m, 1H,pyrrolidine), 3.68 (m, 2H, CH2N), 8.93 (s, 1H, NH) 8

LC/MS: an individual peak at a retention time of 1.9 min, [M + H] ⁺ =222. HPLC under condition C, an individual peak at a retention time of13.8 min. ¹H-NMR (400.13 MHz, DMSO-d₆, δ, m.d., J/Hz): 0.95 (d, 3H, CH₃,J = 6.5 Hz), 2.15 (m, 1H, COCH₂CHCH₂CO); 2.35 (m, 2H, CH ₂C); 2.62 (m,4H, COCH ₂CHCH ₂CO), 3.82 (t, 2H, CH₂ N, J = 7.8 Hz), 6.80 (s, 1H, CCH),7.56 (s, 1H, NCHN)

Example 2 1-(2-(1,3-benzothiazol-2-yl)ethyl)piperidine-2,6-dione(compound 4)

A mixture of 2-(1,3-benzothiazol-2-yl)ethanamide of pentandioic-1,5 acid(22 g; 0.075 mol) and acetic anhydride (23 g; 0.225 mol) were boiled in150 mL of dioxane for 3 hours. The dioxane was removed under vacuum, 200mL of water was added, and the mixture was neutralized with 30% sodiumhydroxide to neutral pH. The precipitated oil was triturated untilcrystals formed. The precipitate was purified by chromatography (SiCO₂60-100 μm, eluent: ethyl acetate-hexane (1:1). LC/MS, an individual peakat a retention time of 2.26 min [M+H]⁺=275. HPLC under condition A,individual peak at a retention time of 9.3 min. ¹H-NMR (400.13 MHz,DMSO-d₆, δ, m.d., J/Hz): 1.85 (quint, 2H, CH₂CH₂CH₂, J=6.8 Hz); 2.59 (t,4H, CH₂CH₂CH₂, J=6.8 Hz); 3.24 (t, 2H, CH₂C, J=7.3 Hz); 4.08 (t, 2H,CH₂N, J=7.3 Hz); 7.43, 7.49 (t, 1H, Ar, J=7.6 Hz); 7.96, 8.04 (d, 1H,Ar, J=7.6 Hz).

The compounds presented in Table 2 were obtained according to theabove-indicated method.

TABLE 3 The number of Structural a compound formula Physical andchemical data 5

LC/MS: an individual peak at a retention time of 0.21 min, [M + H] ⁺ =222. HPLC under condition B, an individual peak at a retention time of20.7 min. ¹H-NMR (400.13 MHz, DMSO- d₆, δ, m.d., J/Hz): 1.82 (quint, 2H,CH₂CH₂CH₂, J = 6.4 Hz), 2.53 (m, 2H, CH₂C), 2.58 (t, 4H, CH₂CH₂CH₂, J =6.4 Hz), 3.57 (s, 3H, NMe), 3.80 (t, 2H, CH₂N, J = 7.8 Hz), 6.85 (s, 1H,CCH), 7.42 (s, 1H, NCHN) 6

LC/MS: an individual peak at a retention time of 1.43 min, [M + H] ⁺ =225. HPLC under condition A, an individual peak at a retention time of31.28 min. ¹H-NMR (400.13 MHz, DMSO- d₆, δ, m.d., J/Hz): 1.82 (quint,2H, CH₂CH₂CH₂, J = 6.5 Hz), 2.58 (t, 4H, CH₂CH₂CH₂, J = 6.5 Hz), 3.12(t, 2H, CH₂C, J = 7.4 Hz), 3.97 (t, 2H, CH₂N, J = 7.4 Hz), 7.58 (d, 1H,SCH, J = 3.2 Hz), 7.70 (d, 1H, NCH, J = 3.2 Hz)

Example 3 Coated tablets, 2 mg, 10 mg, and 100 mg

Composition of a Coated Tablet

Ingredient 2 mg 10 mg 100 mg Active agent: Compound of general 2.00 mg10 mg 100 mg formula I Additives: Microcrystalline cellulose 47.70 mg70.55 mg 95.90 mg Lactose monohydrate 49.00 mg 67.50 mg 99.00 mg Sodiumstarch glycolate 0.50 mg 0.75 mg 1.50 mg Talc 0.40 mg 0.60 mg 1.20 mgMagnesium stearate 0.40 mg 0.60 mg 2.40 mg Tablet core weight 100.00 mg150.00 mg 300.00 mg Film coating 3.00 mg 4.50 mg 9.00 mg Tablet weight103.00 mg 154.50 mg 309.00 mg

Tests of Biological Activity

Materials and Methods

Morphological study of histologic preparations was performed using alight-optical microscope (Leica DM LS, Leica Microsystems, German).Micromorphometric analysis was performed using an ocular micrometerscale mounted in the microscope Leica DM LS. Photomicrographs were madewith a digital photo camera (Leica DC 320).

Mathematical analysis of the obtained results was made using variationstatistics methods with software Statistica 6.0. Data was analyzed withdescriptive statistics: normal distribution of the data was verifiedaccording to Shapiro-Wilk test. As all data fitted normal distribution,the analysis of between-group variance was made by parametric methods,such as Student's t-test. Differences were considered significant ifp<0.05.

An infinite number of examples provided below illustrate the biologicalactivity of the claimed compounds of general formula (I).

Example 4 Evaluation of the Efficiency of the Compounds of GeneralFormula (I) in a Guinea Pig Model of Asthma

Bronchial asthma in guinea pigs was induced by a standard method[Ricciardolo F L, Nijkamp F, De Rose V, Folkerts G. The guinea pig as ananimal model for asthma// Current Drug Targets. 2008 June; 9(6):452-65].The animals were immunized by one parenteral administration of 0.5 mL ofa solution comprising 100 μg/mL ovalbumin (Sigma) and 100 mg/mLaluminium hydroxide. Intact animals received a physiological salinesolution in an amount of 0.5 mL.

On days 29, 30 and 31, hyperactivity of the respiratory airways wasprovoked by inhalation administration of ovalbumin in ascendingconcentrations of 0.1, 0.3, and 0.5 mg/mL on days 1, 2, and 3 of theprovocation, respectively. The inhalation lasted 5 minutes or untilasphyxia symptoms became apparent (fall to one side). On day 32, theanimals received a challenging dose of ovalbumin (1 mg/mL) for 5minutes, while assessing a bronchospastic reaction.

The studied compounds were administered to the animals once daily everyday for 6 or 10 days; the administration was terminated two days beforethe administration of the challenging dose of antigen.

The bronchospastic reaction was assessed by a change in the rate anddepth of the respiratory movements and by the asphyxia symptom such asfall to one side. Spirogram was registered by laboratory experimentalequipment (ADInstruments, Australia), using a base recording station(PowerLab 8sp.) and software LabCart. The registration was made usingairflow sensors for laboratory animals and a spirograph with anintegrated amplifier (ADInstruments).

The bronchoalveolar lavage (BAL) and cardiac cavity blood were derivedfrom the animals 24 hours after administration of the challenging dose.The BAL was collected under anesthesia by lavaging the lungs with 5 mLof a physiological saline solution heated to 37° C. through the trachea,using a syringe dosing device.

The absolute number of cell elements in 1 μL of the lavage (cytosis) wascounted in the bronchoalveolar lavage, using the Goryaev camera. Then,the BAL was centrifuged at 200 g for 10 minutes. The residue was used toprepare swabs that were fixed in methanol and stained according toRomanovsky-Giemsa to count endopulmonary cytogram.

The blood was analyzed with a hemocytometer to determine leykoformula.

The study comprised 3 experiments, wherein the first one was directed tothe evaluation of the effect of claimed compounds 1-6 on the cellcomposition of the BAL, the second and the third experiments were aimedat evaluating the effect of compounds 1 and 2 on the cell composition ofthe BAL, the cell composition of the blood, and on the bronchospasmseverity.

Ten-time administration of compounds 1-6 to guinea pigs at a dose of 14mg/kg reduced the level of cell elements in the BAL from 17.3×10⁹cells/L down to 2.5-6.3×10⁹ cells/L (see Table 4). The studied compoundspreferably resulted in a reduction of the eosinophil count: in thecontrol group, the eosinophil count in the BAL was 7.05×10⁹ cells/L, andin the group receiving the treatment, it was 0.60-1.61×10⁹ cells/L, i.e.less by 91-77%.

TABLE 4 Content of cell elements in BAL in the model of bronchial asthmain guinea pigs (M ± m, n = 9) Dose of compound, Times of Cytosis,Eosinophils, Neutrophils, Monocytes, Group mg/kg administration 10^(9/)π10^(9/)L 10^(9/)L 10^(9/)L Lymphocytes, 10^(9/)L Intact — 1.0 ± 0.1  0.07 ± 0.01 0.16 ± 0.03 0.30 ± 0.05 0.38 ± 0.04 Control — 17.3 ± 1.9*  7.05 ± 0.68* 1.78 ± 0.46 2.70 ± 0.59* 4.92 ± 1.02* Compound 1 14 10 2.5± 0.5&  0.89 ± 0.23*& 0.36 ± 0.09 0.35 ± 0.09* 0.70 ± 0.13* Compound 26.3 ± 1.1*& 1.30 ± 0.29*& 1.05 ± 0.16*& 0.60 ± 0.14* 1.53 ± 0.50*Compound 3 2.9 ± 0.4*& 0.93 ± 0.31*& 0.36 ± 0.08 0.71 ± 0.18& 0.72 ±0.08* Compound 4 3.5 ± 1.1*& 1.25 ± 0.34*& 0.43 ± 0.12 0.85 ± 0.19& 0.81± 0.11* Compound 5 5.9 ± 1.5*& 1.61 ± 0.86*& 1.05 ± 0.33 0.84 ± 0.23*1.35 ± 0.34* Compound 6 4.9 ± 1.2*& 0.60 ± 0.21*& 0.97 ± 0.28* 0.83 ±0.18* 1.03 ± 0.24* Note: *difference from the intact group, according toStudent's t-test, p < 0.05 &difference from the control group Student'st-test, p < 0.05

The administration of the studied compounds at lower doses (0.045, 0.14,and 1.4 mg/kg) and in two regimens (10 and 6 times) showed that thecompounds reduced eosinophilia in BAL within a broad range of doses(0.045 to 14 mg/kg) and in various treatment schemes (see Table 5). Inaddition, the compounds reduced the lymphocyte count in all testeddoses, and the monocyte count in the BAL was also reduced in some doses,which is indicative of the suppression of a local inflammatory reactionin the lung.

TABLE 5 Content of cell elements in BAL in the model of bronchial asthmain guinea pigs (M ± m, n = 8) Dose of compound, Times of Cytosis,Eosinophils, Neutrophils, Monocytes, Lymphocytes, Group mg/kgadministration 10⁹/L 10^(9/)L 10^(9/)L 10^(9/)L 10^(9/)L Experiment No.1 Intact — 0.22 ± 0.01 0.02 ± 0.002 0.06 ± 0.003 0.06 ± 0.006 0.075 ±0.008   Control — 19.3 ± 0.2* 0.96 ± 0.17* 0.18 ± 0.047* 0.54 ± 0.06*0.24 ± 0.03*  Compound 1 0.045 10 12.8 ± 0.2* 0.45 ± 0.07*& 0.19 ± 0.080.52 ± 0.092* 0.10 ± 0.04& 0.14 10.5 ± 0.2*& 0.41 ± 0.10*& 0.07 ± 0.010.43 ± 0.076* 0.13 ± 0.04& 14 14.6 ± 0.3* 0.45 ± 0.12*& 0.15 ± 0.03* 0.7 ± 0.16*  0.15 ± 0.026*& Experiment No. 2 Intact — 0.02 ± 0.003 0.02± 0.003 0.05 ± 0.007 0.05 ± 0.001 0.07 ± 0.01   Control — 0.14 ± 0.02*0.67 ± 0.07* 0.11 ± 0.003 0.49 ± 0.09* 0.16 ± 0.02*  Compound 2 0.14 60.55 ± 0.09*& 0.20 ± 0.03*& 0.03 ± 0.008*& 0.24 ± 0.06*& 0.07 ± 0.02&1.4 0.63 ± 0.09*& 0.18 ± 0.02*& 0.09 ± 0.002 0.29 ± 0.07* 0.06 ± 0.01&14 0.56 ± 0.008*& 0.21 ± 0.06*& 0.07 ± 0.002 0.22 ± 0.05*&  0.07 ±0.009& Note: *difference from the intact group, according to Student'st-test, p < 0.05 &difference from the control group, according toStudent's t-test, p < 0.05

The studied compounds reduced eosinophilia in the blood. In the controlgroup, the eosinophil count in the blood was from 6 to 8 times higherthan in the intact animals. The administration of the studied compoundsallowed for remaining the count at the level of the intact animals (seeTable 6).

TABLE 6 Content of cell elements in the blood in the model of bronchialasthma in guinea pigs (M ± m, n = 8) Dose of Times of Banded SegmentedBaso- Lympho- compound, admin- Cytosis, neutrophils, neutrophils,Eosinophils, phils, Monocytes cytes, Group mg/kg istration 10⁹/L 10⁹/L10⁹/L 10⁹/L 10⁹/L 10⁹/L 10⁹/L Experiment No. 1 Intact — 6.09 ± 0.68  0.08 ± 0.02 2.29 ± 0.35 0.08 ± 0.04 0 ± 0 0.10 ± 0.04 2.66 ± 0.49Control — 12.51 ± 0.68*   0.37 ± 0.11 3.92 ± 0.43 0.46 ± 0.06* 0 ± 00.58 ± 0.16* 7.45 ± 0.62* Compound 1 0.045 10 9.03 ± 1.40&  0.31 ± 0.102.92 ± 0.65 0.19 ± 0.04& 0 ± 0 0.29 ± 0.08 4.80 ± 0.84* 0.14 9.11 ±1.07*& 0.19 ± 0.05& 2.23 ± 0.58& 0.09 ± 0.03& 0 ± 0 0.33 ± 0.06* 5.25 ±0.89* 14 8.90 ± 0.25*& 0.27 ± 0.06* 3.66 ± 0.22* 0.13 ± 0.04& 0 ± 0 0.33± 0.06* 4.45 ± 0.25* Experiment No. 2 Intact 5.89 ± 0.31   0.14 ± 0.042.15 ± 0.26 0.05 ± 0.03 0 ± 0 0.15 ± 0.05 3.41 ± 0.32 Control 13.26 ±0.55*   0.33 ± 0.11 5.34 ± 0.71* 0.39 ± 0.07* 0 ± 0 0.38 ± 0.09* 6.81 ±0.49* Compound 2 0.14 6 7.80 ± 0.77*& 0.18 ± 0.04 3.41 ± 0.41*& 0.09 ±0.03& 0 ± 0 0.18 ± 0.06 3.95 ± 0.54& 1.4 7.70 ± 0.40*& 0.21 ± 0.03 3.53± 0.37*&  0.1 ± 0.03& 0 ± 0 0.18 ± 0.06 3.68 ± 0.25& 14 6.93 ± 0.77& 0.17 ± 0.04  2.9 ± 0.38&  0.1 ± 0.04& 0 ± 0 0.24 ± 0.08 3.51 ± 0.41&Note: *difference from the intact group, according to Student's t-test,p < 0.05 &difference from the control group, according to Student'st-test, p < 0.05

The evaluation of the severity of bronchospasm in the guinea pigs inresponse to inhalation of the challenging dose of ovalbumin showed thatin the model of bronchial asthma, the studied compounds reduced not onlyeosinophilia but also clinical manifestations of the disease. Inparticular, in the control group received placebo, five among eightanimals had severe bronchospasm with acute and subacute phase; suchanimals either were absent in the group of animals received the studiedcompounds or their number was not more than two animals. In turn, thenumber of animals with the normal rate and depth of breathing (withoutbronchospasm) increased from 0-1 in the control group to 4-7 in thegroups received the treatment (see Table 7).

TABLE 7 Severity of bronchospasm in the model of bronchial asthma inguinea pigs The number of animals (n = 8) with moderate with moderatebronchospasm bronchospasm (without acute (without acute phase, withphase, with With severe Dose of subacute phase, subacute phase,bronchospasm compound, Tines of without with breath without breath (inacute and Group mg/kg administration bronchospasm management)management) subacute phase) Experiment No. 1 Intact — 8 0 0 0 Control —0 0 3 5 Compound 1 0.045 10 5 1 2 0 0.14 7 1 0 0 14 5 0 2 1 ExperimentNo. 2 Intact — 8 0 0 0 Control — 1 1 1 5 Compound 2 0.14 6 4 1 1 2 1.4 40 3 1 14 5 1 0 2

The obtained results give reliable evidences that in the experimentalmodels of eosinophilia, in particular of bronchial asthma and the like,the claimed compounds suppress eosinophilia and reduce clinicalmanifestations of the disease.

Example 5 Evaluation of the Efficiency of the Compounds of GeneralFormula (I) in a Model of Sephadex-induced Eosinophilic LungInflammation in Rats

The model of sephadex-induced eosinophilic lung inflammation in rats wasrealized by a standard method [Evaldsson C, Rydén I, Uppugunduri S.Isomaltitol exacerbates neutrophilia but reduces eosinophilia: newinsights into the sephadex model of lung inflammation//Int Arch AllergyImmunol. 2011; 154(4):286-94]. Sephadex G-200 (Pharmacia, Sweden) wasadministered one time by inhalation to male Wistar rats at a dose of 5mg/kg. The studied compounds were administered to the animals by theintragastric route four times: 24 hours and 1 hour before and 24 hoursand 45 hours after administration of sephadex. The referencepreparation, budesonide, was administered according to the same scheme,by inhalation at a dose of 0.5 mg/kg. Bronchoalveolar lavage was taken48 hours after the inhalation of sephadex, and the total lymphocytecount and leukocyte formula in the lavage were determined. The number ofrats in a group was 7 to 10.

The analysis of the BAL showed that one time administration of sephadexG-200 by inhalation caused an apparent flow of leukocytes into the lung.The content of all cell types was increased in the control grouprelative to the intact animals; however, the maximum increase wasregistered for eosinophils (see Tables 8-9).

The intragastric administration of the compounds of formula (I) to therats reduced the eosinophil count in the BAL by several times. Theclaimed compounds exhibited activity within a broad range of the testeddoses.

TABLE 8 Content of cell elements in BAL in the model of sephadex-inducedeosinophilic lung inflammation in rats (M ± m, n = 10) Content of cellelements in 1 μL of BAL Group Leukocytes Neutrophils EosinophilsMacrophages Lymphocytes Intact  865 ± 78  35 ± 9  13 ± 4  770 ± 66  46 ±12 Control 2785 ± 152* 711 ± 158* 444 ± 45* 1469 ± 197* 161 ± 44*Compound 1 (0.06 mg/kg) 2500 ± 307* 837 ± 231* 118 ± 27*& 1417 ± 131*128 ± 48 Compound 1 (0.18 mg/kg) 2665 ± 455* 597 ± 168* 174 ± 64*& 1661± 260* 143 ± 54 Compound 1 (0.54 mg/kg) 2120 ± 218*& 352 ± 135* 126 ±43*& 1446 ± 113* 196 ± 63* Compound 1 (1.8 mg/kg) 1915 ± 250*& 451 ±149* 129 ± 44*& 1214 ± 130* 122 ± 49 Compound 1 (5.4 mg/kg) 2340 ± 322*492 ± 129* 152 ± 56*& 1525 ± 199* 170 ± 45* Compound 1 (18 mg/kg) 2135 ±205*& 297 ± 89*&  69 ± 22*& 1601 ± 134* 168 ± 44* Budesonide (0.5 mg/kg)1805 ± 318*& 334 ± 119* 204 ± 84*& 1230 ± 205  37 ± 10& Note:*difference from the intact group, according to Student's t-test, p <0.05 &difference from the control group, according to Student's t-test,p < 0.05

TABLE 9 Content of cell elements in BAL in the model of sephadex-inducedeosinophilic lung inflammation in rats (M ± m, n = 7) Content of cellelements in 1 μL of BAL Group Leukocytes Neutrophils EosinophilsMacrophages Lymphocytes Intact 2471 ± 611 150 ± 63  39 ± 31 1532 ± 225 31 ± 26 Control 5207 ± 814* 785 ± 163* 630 ± 104* 3471 ± 412* 275 ± 103Compound 5 (1.8 mg/kg) 2064 ± 257& 385 ± 57* 147 ± 44& 1576 ± 93&  66 ±26 Compound 5 (18 mg/kg) 2189 ± 222& 605 ± 124  34 ± 15& 1331 ± 66 & 121± 48 Compound 7 (1.8 mg/kg) 2242 ± 289& 389 ± 188  28 ± 16&  913 ± 235& 16 ± 10 Compound 7 (18 mg/kg) 1725 ± 345& 136 ± 50& 164 ± 62 &  909 ±299&  29 ± 17 Compound 8 (1.8 mg/kg) 2796 ± 333 & 303 ± 104& 202 ± 53*&1860 ± 204&  82 ± 44 Compound 8 (18 mg/kg) 4250 ± 576 583 ± 180 318 ±65*& 2050 ± 462&  64 ± 45 Note: *difference from the intact group,according to Student's t-test, p < 0.05 &difference from the controlgroup, according to Student's t-test, p < 0.05

Example 6 Evaluation of the Efficiency of the Compounds of GeneralFormula (I) in a Model of Leukotriene-induced Eosinophilic LungInflammation in Guinea Pigs

The model of leukotriene-induced eosinophilic lung inflammation inguinea pigs was realized by a standard method [Underwood DC1, Osborn RR, Newsholme S J, Torphy T J, Hay D W. Persistent airway eosinophiliaafter leukotriene (LT) D4 administration in the guinea pig: modulationby the LTD4 receptor antagonist, pranlukast, or an interleukin-5monoclonal antibody// Am J Respir Crit Care Med. 1996 October; 154(4 Pt1):850-7]. A solution of leukotriene D4 (LTD4, Cayman Chemical, USA) ata concentration of 10 mg/kg (a flow rate of 250 mL/hr) was inhaled tomale guinea pigs (250-300 g) under conditions of a double-chamberplethysmograph (Emka Technologies, France) for one minute. The studiedcompound was administered to the animals by the intragastric route fourtimes: 24 hours and 1 hour before and 24 hours and 45 hours after theinhalation of LTD4. The reference preparation, montelukast (0.8 mg/kg),was administered one time by the intragastric route one hour before theinhalation of LTD4. Bronchoalveolar lavage was taken 48 hours after theinhalation of LTD4, and the total lymphocyte count and leukocyte formulawere determined in the lavage. The number of guinea pigs in a group was8.

The analysis of BAL showed that one time administration of leukotrieneD4 to guinea pig by inhalation caused an apparent flow of neutrophils,eosinophils, and monocytes/macrophages into the lungs. The most evidentincrease in the cell count (25 times) was observed for eosinophils (seeTable 10).

The intragastric administration of the studied compound to the guineapigs reduced the eosinophil count in the BAL by 2.1-3.4 times. Thecompounds had an effect within a broad range of doses (0.14-14 mg/kg).The comparative analysis of the efficiency of the claimed compound andmontelukast showed that the effects of the claimed compound and theleukotriene receptor agonist were comparable by strength.

TABLE 10 Content of cell elements in BAL in the model ofleukotriene-induced eosinophilic lung inflammation in guinea pigs (M ±m, n = 8) Content of cell elements in 1 μL of BAL Group LeukocytesNeutrophils Eosinophils Macrophages Lymphocytes Intact   556 ± 83  21 ±4  79 ± 26  357 ± 55 100 ± 32 Control  5788 ± 1269* 303 ± 66* 1966 ±391* 3392 ± 895* 126 ± 35 Compound 1 (0.14 mg/kg)  2638 ± 463*& 249 ± 50 916 ± 144*& 1360 ± 323* 112 ± 27 Compound 1 (1.4 mg/kg)  3413 ± 1022*279 ± 97*  856 ± 288*& 2134 ± 705* 144 ± 89 Compound 1 (14 mg/kg)  2250± 373*& 155 ± 36*  580 ± 124*& 1444 ± 258*  71 ± 28 Montelukast (0.8mg/kg) 24065 ± 415*& 173 ± 41*  618 ± 116*& 1519 ± 263*  96 ± 39 Note:*difference from the intact group, according to Student's t-test, p <0.05 &difference from the control group, according to Student's t-test,p < 0.05

Example 7 Evaluation of the Efficiency of the Compounds of GeneralFormula (I) in a Model of Allergic Rhinitis in Guinea Pigs

The model of allergic rhinitis in guinea pigs was realized by a standardmethod [Vishnu N. Thakare, M. M. Osama, Suresh R. Naik. Therapeuticpotential of curcumin in experimentally induced allergic rhinitis inguinea pigs // Int Immunopharmacol. 2013 September; 17(1):18-25].

Guinea pigs (250-300 g) were immunized with 4-time (on days 0, 7, 14,and 21) intragastric administration of ovalbumin (100 μg/pig) andaluminium hydroxide (5 mg/pig), both of which were diluted and suspendedin a physiological saline solution. On day of the study, a solution ofovalbumin (60 mg/mL) was administered intranasally to each nostril ofthe animals at a dose 20 μL. On day 35, the animals receivedsubcutaneously an ovalbumin solution (200 μg/mL, 25 μL); the animal backwas previously shaved in the site of administration. Swelling andredness at the injection site served as a support of sensibilisation. Onday 42 of the study, an ovalbumin solution (60 mg/mL, 20 μL/nostril) wasadministered intranasally. A group of pseudoimmunized animals was formedto control the formation of exactly allergic inflammation: on days 0, 7,14, and 21, the pigs received a solution of aluminium hydroxide (5mg/pig), and on days 28 and 35, they received a physiological salinesolution, and on day 42, a solution of ovalbumin (60 mg/mL, 20μL/nostril).

The studied compounds (14 mg/kg) were administered 3 times by theintragastric route: 48, 24, and 1 hour before the last administration ofovalbumin. The reference preparation, dexamethasone, was administeredone time by the intragastric route, 3 hours before the last intranasaladministration of ovalbumin.

Clinical manifestations, such as the number of sneezes and nosescratchings, were registered for 2 hours after the last administrationof ovalbumin. Nasal lavage was taken 24 hours after the lastadministration of ovalbumin, and in this lavage, the total lymphocytecount and leukocyte formula were determined. The number of guinea pigsin a group was 8.

The analysis of the nasal lavage showed that allergic rhinitis isaccompanied by an apparent flow of leukocytes into the nasal cavity. Themaximum increase was registered for eosinophils (Table 11).

Three-time administration of the compounds of general formula (I) to theguinea pigs reduced the eosinophil count in the nasal lavage to thelevel observed in the pseudoimmunized animals. The effects of theclaimed compound and dexamethasone were comparable by strength.

TABLE 11 Content of cell elements in the nasal lavage of in guinea pigsin the model of allergic rhinitis (M ± m, n = 8) Content of cellelements in 1 μL of nasal lavage Group Leukocytes NeutrophilsEosinophils Macrophages Lymphocytes Pseudoimmunization 1371 ± 181 424 ±30  267 ± 38 712 ± 125 8 ± 6 Control 3029 ± 286* 753 ± 121* 1265 ± 226*439 ± 132 8 ± 8 Compound 1 (14 mg/kg) 1300 ± 254& 243 ± 58*&  521 ± 129&405 ± 130 6 ± 6 Compound 5 (14 mg/kg) 1071 ± 233& 344 ± 50&  575 ± 143&198 ± 54* 9 ± 7 Dexamethasone (5 1186 ± 142& 328 ± 47&  365 ± 90& 296 ±71* 5 ± 3 mg/kg) Note: *difference from the intact group, according toStudent's t-test, p < 0.05 &difference from the control group, accordingto Student's t-test, p < 0.05

The registration of clinical manifestations of allergic rhinitis for 2hours after the last intranasal administration of ovalbumin showed anapparent increase in the number of sneezes and nose scratchings in theexperimental animals, which was indicative of the correctness of therealized model of allergic rhinitis. The therapy with the compounds ofgeneral formula (I) reduced the clinical manifestations of rhinitis totheir level observed in the pseudoimmunized animals. The referencepreparation had a similar effect (see Table 12).

TABLE 12 Clinical manifestations of allergic rhinitis in guinea pigs inthe experimental model (M ± m, n = 8) Number of Number of nose Groupsneezes/2 hours scratchings/2 hours Pseudoimmunization  5.3 ± 1.2  9.7 ±1.3 Control 16.3 ± 2.6* 45.3 ± 5.2* Compound 1 (14 mg/kg)  5.9 ± 1.1 &13.9 ± 2.6 & Compound 5 (14 mg/kg)  7.1 ± 1.3 & 19.1 ± 4.9 &Dexamethasone (5 mg/kg)  7.9 ± 0.8 & 16.7 ± 1.8*& Notes: *differencefrom the intact group, according to Student's t-test, p < 0.05 &difference from the control group, according to Student's t-test, p <0.05

Example 8 Evaluation of the Efficiency of the Compounds of GeneralFormula (I) in a Model of Allopathic Dermatitis in Mice

The model of allopathic dermatitis was realized by a standard method[Mechanism of dinitrochlorobenzene-induced dermatitis in mice: role ofspecific antibodies in pathogenesis//PLoS One. 2009; 4(11)].

On days 0 and 12 of the study, 100 μL of a 2% solution of1-chloro-2,4-dinitrobenzene (DNCB, Sigma-Aldrich, US) in 95% ethanol wasapplied to shaved sites in the backs of male Balb/c mice. On day 17 ofthe study, to the right “studied” ear of the animals, 20 μL of thealcohol solution of 2% DNCB was applied two times with a one-hourinterval. The studied compound and the reference preparation,dexamethasone, were administered by the intragastric route once daily ondays 8-17 of the study.

On day 18 of the study, the animals were euthanized in a CO₂ camera. Theweights of the “studied” and “control” ears were measured. A responseindex (RI) expressed in percentage of a difference in the weights of the“studied” and “control” ears was calculated.

The study showed that the compounds of general formula (I) reduced theresponse index in the experimental model of atopic dermatitis. Theeffects of the claimed compound and of the steroid preparation,dexamethasone, were comparable by strength (see Table 13).

TABLE 13 Response index in atopic dermatitis in mice (M ± m, n = 12)Group Response index (%) Intact −0.49 ± 0.68 Control  93.8 ± 5.4*Compound 1 (0.3 mg/kg)  74.4 ± 7.3*& Compound 1 (3 mg/kg)  69.5 ± 8.2*&Compound 1 (30 mg/kg)  69.1 ± 8*& Dexamethasone (10 mg/kg)  65.6 ± 8*&Notes: *difference from the intact group, according to Student's t-test,p < 0.05 &difference from the control group, according to Student'st-test, p < 0.05

The obtained results give grounds for a conclusion that in theexperimental models of eosinophilia, in particular sephadex-inducedeosinophilic lung inflammation in rats, leukotriene-induced eosinophiliclung inflammation in guinea pigs, allergic rhinitis and asthma in guineapigs, atopic dermatitis in mice, and the like, the compounds of generalformula (I) significantly reduce eosinophilia.

What is claimed is:
 1. A method of treating an eosinophilic disease, themethod comprising administering to a patient of an effective amount of acompound of the formula

or a pharmaceutically acceptable salt thereof, wherein the eosinophilicdisease is selected from the group consisting of bronchial asthma,allergic rhinitis, polypous rhinosinusopathies, atopic dermatitis, andeosinophilic esophagitis, and wherein eosinophils in the patient arereduced.
 2. The method of claim 1, wherein the pharmaceuticallyacceptable salt is a salt of an organic acid, a salt of an inorganicacid, or a salt of an amino acid.
 3. The method of claim 2, wherein thesalt of the organic acid is a formate salt, an acetate salt, a maleatesalt, a tartrate salt, a methanesulfonate salt, a benzenesulfonate salt,or a toluenesulfonate salt.
 4. The method of claim 2, wherein the saltof the inorganic acid is a hydrochloride salt, a hydrobromide salt, asulfate salt, or a phosphate salt.
 5. The method of claim 2, wherein thesalt of the amino acid is an aspartic acid salt or a glutamic acid salt.6. The method of claim 1, wherein the pharmaceutically acceptable saltis a chlorohydrate or an acetate.
 7. The method of claim 1, wherein theadministering of the compound or pharmaceutically acceptable saltthereof is carried out by an administration route selected from thegroup consisting of oral, topical, parenteral, intranasal, sublingual,inhalational, and rectal.
 8. The method of claim 7, wherein theadministration route is parenteral and the compound is administeredsubcutaneously, intravenously, intramuscularly, by injection, or byinfusion.
 9. The method of claim 1, wherein the administering of thecompound or pharmaceutically acceptable salt thereof is carried out at adose of from about 0.1 to about 30 mg/kg of body weight of the patient.10. The method of claim 9, wherein the dose is from about 0.25 to about10 mg/kg of body weight of the patient.
 11. The method of claim 9,wherein the dose is administered one or more times per day.
 12. Themethod of claim 11, wherein the dose is administered once daily.
 13. Themethod of claim 1, wherein the compound or pharmaceutically acceptablesalt thereof is administered in a pharmaceutical composition, furthercomprising a pharmaceutically acceptable carrier.
 14. The method ofclaim 13, wherein the pharmaceutical composition is in a form selectedfrom the group consisting of a solution, a tablet, a pill, a capsule, adragee, a suppository, an emulsion, a suspension, an ointment, and agel.
 15. The method of claim 14, wherein the pharmaceutical compositionis in the form of a pill, coated with a layer that is resistant togastric acid.
 16. The method of claim 13, wherein the pharmaceuticallyacceptable carrier selected from the group consisting of glucose,lactose, sucrose, mannitol, sorbitol, a cellulose derivative, and acalcium phosphate.
 17. The method of claim 13, wherein thepharmaceutical composition further comprises a binder.